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Retention of lysozyme activity by physical immobilization in nanocellulose aerogels and antibacterial effects

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Abstract

Aerogels prepared from aqueous dispersions of anionic and cationic cellulose nanofibrils (CNFs) were investigated as solid supports for enzymes and silver nanoparticles and to elicit a sustained antibacterial effect. The imparted stabilization in dry conditions was studied with aerogels that were cast after mixing the enzymes with CNFs followed by dehydration (freeze-drying). The activity of lysozyme immobilized in the given CNF system was analyzed upon storage in liquid and air media. In contrast with aqueous solutions of free, unbound enzyme, which lost activity after the first day, the enzyme immobilized physically in unmodified and cationic CNF presented better stability (activity for a longer time). However, the enzyme activity was reduced in the case of anionic CNF, which was prepared by TEMPO-mediated oxidation (TO-CNF). Both humidity and temperature reduced the stability of the enzyme immobilized in the respective CNF aerogel. The antibacterial activity of CNF aerogels carrying lysozyme was also tested against gram-negative and gram-positive bacteria. The results were compared with those obtained from CNF systems loaded with silver nanoparticles (AgNP) after in situ synthesis via UV reduction. Storage in cold or dry conditions preserved the activity and antibacterial performance of enzyme-loaded CNF aerogels. As expected, the lysozyme-containing aerogels showed lower inhibition than the AgNP-containing aerogel. In this latter case, the antibacterial activity depended on the concentration and size of the nanoparticles. Compared to unmodified CNF and TO-CNF, the aerogels prepared with cationic CNF, loaded with either lysozyme or AgNPs, showed remarkably better antibacterial activity. Similar experiments were conducted with horseradish peroxidase, which confirmed, to different degrees, the observations derived from the lysozyme systems. Overall, the results indicate that non-toxic and biodegradable CNF is a suitable support for bio-active materials and is effective in protecting and retaining enzymatic and antibacterial activities.

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Acknowledgments

The authors are grateful for funding support by the Academy of Finland (AF) through Project 295007, Nanocellulose Cyber-Physical Microsystems (ICT 2023: Advanced microsystems) as well as AF’s Centres of Excellence Programme (2014–2019), under Project 264677 “Molecular Engineering of Biosynthetic Hybrid Materials Research” (HYBER). This work made use of the Aalto University Nano microscopy Center.

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Authors and Affiliations

  1. Department of Bioproducts and Biosystems, School of Chemical Engineering, Aalto University Aalto, Espoo, Finland

    Khan M. A. Uddin, Hannes Orelma, Pezhman Mohammadi, Maryam Borghei, Janne Laine, Markus Linder & Orlando J. Rojas

  2. VTT Technical Research Centre of Finland, High Performance Fibre Products, Espoo, Finland

    Hannes Orelma

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  1. Khan M. A. Uddin
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Correspondence to Hannes Orelma or Orlando J. Rojas.

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Uddin, K.M.A., Orelma, H., Mohammadi, P. et al. Retention of lysozyme activity by physical immobilization in nanocellulose aerogels and antibacterial effects. Cellulose 24, 2837–2848 (2017). https://doi.org/10.1007/s10570-017-1311-0

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